Discovery of three novel G-protein-coupled receptor genes.
We report here the molecular cloning, tissue distribution, and chromosomal localization of novel genes encoding G-protein-coupled receptors (GPCRs). A search of a mouse database of expressed sequence tags revealed an EST partially encoding a GPCR, which was used to screen a mouse genomic library to obtain the translational open reading frame (ORF). The resultant clone, GPR27, contained an intronless ORF, encoding a receptor of 379 amino acids. In an alternate strategy, human genomic DNA was subjected to polymerase chain reaction (PCR) amplification, using degenerate oligonucleotides based on GPR1. Two PCR products partially encoding GPCRs were isolated and used to screen a genomic library to obtain the translational ORF. One of the resultant clones, GPR30, contained an intronless ORF encoding a receptor of 375 amino acids. The other clone, GPR35, also contained an intronless ORF encoding a receptor of 309 amino acids. Transcripts corresponding to GPR27 and GPR30 were detected in several areas of human and rat CNS, While GPR35 expression was detected only in the rat intestine. Through fluorescence in situ hybridization analysis the gene encoding GPR30 was localized to chromosome 7p22 and GPR35 to chromosome 2q37.3.
O'Dowd
B F
BF
Department of Pharmacology, University of Toronto, Ontario, Canada. brian.odowd@utoronto.ca
Nguyen
T
T
Marchese
A
A
Cheng
R
R
Lynch
K R
KR
Heng
H H
HH
Kolakowski
L F
LF
George
S R
SR
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000235:genetics; D002889:Chromosomes, Human, Pair 2; D002897:Chromosomes, Human, Pair 7; D003001:Cloning, Molecular; D019204:GTP-Binding Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009928:Organ Specificity / Q000235:genetics; D051381:Rats; D044087:Receptor, Anaphylatoxin C5a; D011956:Receptors, Cell Surface / Q000032:analysis / Q000235:genetics*; D011951:Receptors, Complement / Q000235:genetics; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D017386:Sequence Homology, Amino Acid
D015703:Antigens, CD; C109584:GPER1 protein, human; C109334:GPR1 protein, S cerevisiae; C111379:GPR35 protein, human; C111343:Gpr27 protein, mouse; D044087:Receptor, Anaphylatoxin C5a; D011956:Receptors, Cell Surface; D011951:Receptors, Complement; D011960:Receptors, Estrogen; D043562:Receptors, G-Protein-Coupled; D029701:Saccharomyces cerevisiae Proteins; D019204:GTP-Binding Proteins